|
|
Glutamine and Lysine Determination
Two Isobaric Amino Acids in Saliva
Click here to view
printable Application Sheet
|
Notes: Adapted from: "Analysis of Hydrophilic Metabolites in Physiological Fluids by HPLC-MS using
a Silica Hydride-Based Stationary Phase", J.J. Pesek, M.T. Matyska, J.A. Loo, S.M. Fischer, T.R.
Sana, J. Sep. Sci., 32 (2009) 2200-2208.
|
|
Method Conditions
| Column |
Cogent Diamond Hydride™, 4µm, 100A |
| Catalog No. |
70000-15P-2 |
| Dimensions |
2.1 x 150 mm |
| Solvents |
A: DI water + 10 mM ammonium acetate
B: 98% Acetonitrile + 2% (10 mM ammonium acetate) |
| Gradient |
| Time (min) |
%B |
| 0.0 |
100 |
| 14.00 |
60 |
| 14.10 |
5 |
|
| Post time |
5 min, column temperature 25°C |
| Flow rate |
0.4 mL/min. t0 = 0.8 min |
| Injection |
1 µL |
| Samples |
1. D,L - glutamine (M+H)+, 147.0764 m/z
2. D,L - lysine (M+H)+, 147.1134 m/z
|
| Detection |
ESI – pos - Agilent 6210 MSD TOF mass spectrometer. |
Discussion
The chromatogram above shows an interesting aspect of analyzing real patient samples for a pair of metabolites:
glutamine and lysine. Figure A represents 7 samples taken from patients with cancer; Figure C shows injections of
samples from 7 patients with pancreatitis while Figure B represents 7 samples from a control group of healthy
people. In the samples studied generally the lysine peak intensity was approximately equal to or greater than the
glutamine peak intensity in cancer patients while in the normal patients the lysine peak was significantly lower
in intensity than the glutamine peak. Please note the reproducibility of the analysis (retention times) for all
samples, despite the variability in the concentration level of the two amino acids in saliva.
|
